基因工程菌株BLG8900遗传稳定性.pdfVIP

98 2007 2 20 2 Ch in J B io log icals February 2007, V o1. 20 No. 2 Q754 A ( 2007 ) 0209803 BLG8900 1, 2 3 4 5 2 2 金元昌 磨美兰 苏晓艳 李景鹏 向育君 李会东 BLG8900 (), pYC2 BLG 8900[ pYC2 BL21( D E3) ] 102550 100 , BamH /EcoR , 100 GnRH /TR S 102550 100 PTG , GSTG nRH /TR S SD SPAG E W estern blot G nRH pYC2 BL2 1 ( DE3) ; ; G enetic Stability of RecombinantE. coli BLG8900 Strain J N Yuanchang , MO M e ilan, SU X iaoyan, et a l ( C olleg e of L fi e Science and Technology, Guangx i Un ivers ity, N ann ing 530004, Ch ina ) Ab tract Objective T o study the g enetic stab ility of recom binant E. co li BLG 8900 stra in. M ethod Subcu lture recomb i nant E. co li BLG 8900 stra in, a BL 21( DE3) stra in contain ing plasm id pYC2, for different passages and test fo r the losing rate of plas m id. R e u lt A fte r subcu lture fo r 10, 25, 50 and 100 passages, the losing rate of p lasm id pY C2 in recomb inan tE. co li BLG8900 strain w ere 1% , 1% , 2% and 2% respec tive ly, and the restr iction maps of bacte riaw ere identical. The G nRH /TR S sequence of plasm id in re comb inant E. co li bacte ria passag e 100 w as identica l to that in orig inal recom b inant E. co li. Both the expression lev els and SD SPAGE profiles of fusion pro tein G STG nRH /TR S in recomb inan tE. co li BLG 8900 strain of various passages show ed no significant d ifference. T he sequence of fus ion g ene in the BLG8900 stra in of passag e 100 w as comp le tely identica l to that of passage 0. W estern b lo t proved G nRH antigen ic spec ific ity of expressed product in BLG 8900 stra in of various passages. Con clu ion The recomb inantE. coli BLG 8900 stra in containing plasm id pYC2 show ed good gene tic stability. K ey word G onadotropin re leas ing horm one( GnRH ); R